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The energetic cost of protein synthesis in isolated hepatocytes of rainbow trout (Oncorhynchus mykiss)

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Summary

To establish the energetic cost of protein synthesis, isolated trout hepatocytes were used to measure protein synthesis and respiration simultaneously at a variety of temperatures. The presence of bovine serum albumin was essential for the viability of isolated hepatocytes during isolation, but, in order to measure protein synthesis rates, oxygen consumption rates and RNA-to-protein ratios, BSA had to be washed from the cells. Isolated hepatocytes were found to be capable of protein synthesis and oxygen consumption at constant rates over a wide range of oxygen tension. Cycloheximide was used to inhibit protein synthesis. Isolated hepatocytes used on average 79.7±9.5% of their total oxygen consumption on cycloheximide-sensitive protein synthesis and 2.8±2.8% on maintaining ouabain-sensitive Na+/K+-ATPase activity. The energetic cost of protein synthesis in terms of moles of adenosine triphosphate per gram of protein synthesis decreased with increasing rates of protein synthesis at higher temperatures. It is suggested that the energetic cost consists of a fixed (independent of synthesis rate) and a variable component (dependent on synthesis rate).

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Abbreviations

BSA:

bovine serum albumin

dpm:

disintegrations per min

k s :

fractional rate of protein synthesis

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethane sulphonic acid

PHE:

phenylalanine; PO2 oxygen tension

PCA:

perchloric acid

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Author notes

  1. Marinus C. Pannevis

    Present address: Waltham Centre for Pet Nutrition, Freeby Lane, Waltham on the Wolds, LE14 4RT, Melton Mowbray, UK

Authors and Affiliations

  1. Department of Zoology, University of Aberdeen, Tillydrone Avenue, AB9 2TN, Aberdeen, UK

    Marinus C. Pannevis & Dominic F. Houlihan

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  1. Marinus C. Pannevis
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  2. Dominic F. Houlihan
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Pannevis, M.C., Houlihan, D.F. The energetic cost of protein synthesis in isolated hepatocytes of rainbow trout (Oncorhynchus mykiss). J Comp Physiol B 162, 393–400 (1992). https://doi.org/10.1007/BF00258960

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